Figure 1.

(A) Schematic of suggested approach for DNA part library assembly and characterization in vitro. (i) Generation of a DNA part library is achieved by PCR amplification of DNA regulatory elements upstream of a GFP reporter gene. (ii) These PCR-amplified linear DNA products are circularized by intramolecular ligation to form closed-circular DNA. (iii) Closed-circular DNA products are used as DNA templates for characterization in cell-free systems by fluorescence measurements. Characterization data can be used to annotate DNA part libraries, and closed-circular DNA is stored. (B) Example of current methodology for DNA part assembly and characterization in cell-free systems.